Oral and Poster Abstracts

A 5-month-old calf with a chronic diarrhea and bloat and obvious
abdominal pain was referred to the large Animal Teaching and
Research Hospital. On physical examination the calf was lethargic and
had a rectal temperature of 39.8 ˚C, respiratory rate 44 breaths per
minutes, heart rate 100 beats per minutes and mild tachycardia. The
calf had pale mucosal membranes and clinical dehydration. The
peritoneal fluid examination showed an exudate with high protein level
and degenerated neutrophil which was positive for Salmonella dublin.
The fecal samples were negative for parasites and Mycobacterium
paratuberculosis, but positive for Salmonella dublin. The leukogram
showed significant leukocytosis, lymphocytosis and monocytosis
accompanying by neutropenia and a shift to left. A degenerative
anemia was diagnosed in the calf which could be a ambiguous
prognosis. The animal died despite of relevant fluid and antibiotic
therapy, and a chronic extensive peritonitis was found in the
postmortem examination
Key words: calf, chronic extensive peritonitis, diarrhea, leukogram
647 Molecular Survey on Occurrence of Mycoplasma Mycoides
Subspecies Mycoides L.C and S.C in Pulmonary Infection of
Iranian Cattle, Goats and Sheep
N. Atyabi, Z. Nikusefat, SA. Pourbakhsh
Faculty of Veterinary Medicine, University of Tehran, Clinical
Science, Tehran, Iran
This survey described microbiological and molecular study for detection
Mycoplasma mycoides subspecies mycoides (L.C and S.C) in pneumonic
lungs of Iranian sheep, goats and cattle. Suspected samples were
investigated based on color, consistency and appearance of the cut
surface. Total 180 lung samples were collected from slaughter house
located in West of Iran, Kermanshah during 2006-2008 and transferred
near ice packs to Razi Research Institute. Gross lesions showed
hepatization with grey and white lesions (consolidation) and motley
appearance with or without fibrin. Minced lung tissues were inoculated to
PPLO broth agar (Diffco) with application acetatecellulose filter (0.45-2.5
micrometer). Samples were in 6.8% carried to Co2 incubator. After 10-14
days the yellow tubes were subcultured to PPLO plate. After while,
characteristic colony was observed. DNA extractions were based on
phenol method and DNA extracted isolates were preserved in 50%
glycerol at -20˚C.DNA extract of all samples were subjected to generic
and species specific PCR based on 16 S rRNA with different set of
primers and cycles. The visualized amplicon consisted of 573 bp.40
samples from total 180 lung extract were positive for genus mycoplasma
(22.2%),while only 30 samples were positive in culture(16.66%) .There
were no significance difference in sex and age between affected
animals.(P>0.05) The highest percentage of infection was observed
during December (32%) and the lowest was in June (2.5%).There were no
evidence of Mycoplasma mycoides mycoides (S.C ) and (L.C) by species
specific primer based on CAP-21 PCR in infected ruminants. However,
the present work was carried out to study the incidence of two strains of
mycoid cluster in Iranian ruminants. Further investigation should be
conducted to profile all mycoplasma spp in pulmonary infections.
Key words: Mycoplasma mycoides, sub. mycoides (L.C and S.C),
ruminant, Iran
648 Application of PCR and Hematological Findings in
Replacement Dairy Heifers with ELISA Seropositive to
Bovine Leukosis Virus Infection
T. Rukkwamsuk, S. Panneum
Faculty of Veterinary Medicine, Kasetsart University, Large Animal
and Wildlife Clinical Science, Nakhon-Pathom, Thailand
Bovine leukosis virus infection was studied in 171 replacement dairy
heifers. Blood samples were collected for determination of
hematological parameters, detection of bovine leukosis virus using
PCR technique, and determination of antibody against bovine leukosis
virus infection using indirect ELISA (IDEXX HerdCheck Anti-BLV).
Blood samples from seropositive and seronegative heifers were
randomly selected to detect bovine leukosis virus using PCR technique.
Results revealed that seroprevalence of bovine leukosis virus infection
in this group of heifers was 19.3% (33 heifers were seropositive and
138 heifers were seronegative). Most hematological parameters did not
differ between seropositive and seronegative heifers. Average total
white cell counts were higher for seropositive heifers than for
seronegative heifers (P = 0.08). Regarding PCR results, bovine
leukosis virus was not detected from some seropositive heifers but
could be detected from some seronegative heifers. However, results
obtained from indirect ELISA and from PCR technique had a moderate
agreement (Kappa value = 0.60). In conclusion, seroprevalence of
bovine leukosis virus infection in replacement dairy heifers was
relatively high. Increased total white cell count seemed to be related to
bovine leukosis virus infection. Although agreement of serological test
using ELISA and virus detection using PCR was moderate, results of
these tests were still contradicted in some heifers. Therefore, further
study on increasing sensitivity and specificity of diagnostic tests is
required.
Key words: Bovine Leukosis Virus, hematology, replacement heifer
649 Bovine Leptospirosis in Sardinia: Isolation and Evaluation of
Molecular Methods
M. Ponti, G. Sanna, G. Carboni, G. Canu, M. Manca, M. Noworol, B.
Palmas, E. Marongiu, C. Patta
Istituto Zooprofilattico Sperimentale Della Sardegna, Sassari, Italy
Previous studies carried out in Sardinia have shown that Leptospirosis
affects both humans and farm animals and that Leptospira serovar
pomona is responsible for the icterus-haemorrhagic syndrome in
calves. In this study, the microscopic agglutination test (MAT) was
used to determine the serological titers of the serum samples collected
from cattle. Even though definitive diagnosis is provided by culture of
pathogenic Leptospira, several molecular approaches were also
evaluated, aiming at optimizing the detection of bacteria in bovine
samples of urine and blood. Specimens were collected from two dairy
cattle herds in the north of Sardinia. Farm A consisted of 580 subjects,
out of which 260 were lactating cows, 280 were freerange heifers and
40 were calves. Farm B included 120 animals, out of which 45 were
lactating cows and the remaining ones were heifers and calves. Sera
were examined by MAT. Urine, blood and tissue suspension were
inoculated in semi-solid medium for cultivation and tested by PCR.
The Leptospires strains isolated were identified by using monoclonal
antibodies. Several primer pairs for detection of leptospiral DNA were
tested: one set amplified a fragment of the 16S rRNA, another set was
complementary to a portion of Lig protein gene (Lig 1/Lig 2) and a
third one used G1/G2 and B64I/B64II. Then a protocol of real-time
PCR using a Taqman Probe was standardized and compared with the
three protocols previously described. In farm A the seroprevalence was
of 60%, while in farm B it was about 40%. Leptospires were isolated
from 8 and 3 urine samples in farms A and B, respectively. All strains
were identified as Leptospira serovar pomona. Leptospires were not
isolated from blood and fetuses samples. All the molecular methods
used amplified leptospiral DNA from all 15 serovars of our Leptospires
panel, and also from blood, urine and fetuses tissues. Primers derived
from rRNA gene sequence were the least specific, and none of the
molecular methods tested was 100% sensitive. Furthermore, the main
limitation of these PCR-based assays was their inability to identify the
infecting serovar. Real-time PCR assay, which successfully detected
leptospiral DNA, is rapid and specific but more expensive than
conventional PCR. A combination of two detection methods (PCR and
culture) is the most sensitive approach for early diagnosis of
Leptospirosis.
Key words: leptospirosis, bovine, PCR
650 Serological Surveillance of Contagious Bovine
Pleuoropneumonia between 2005 and 2007 Years in Romania
R. Radulescu, G. Petriceanu, A. Ragalie, E. Gutu
Institute for Diagnosis and Animal Health, Immunology, Bucharest,
Romania
The reference method recommended by OIE for serological
surveillance of Contagious Bovine Pleuropneumonia (CBPP) is the
Complement Fixation Test (CFT). This method had been used in the
past for eradication of CBPP in many countries. However, CFT have
some disadvantages, mainly the production and standardization of
specific antigen, providing some false positive reaction due to the
mycoplasmas cross-reactivity from M. mycoides cluster. In Romania
between 2005 and 2007, years have been examined for CBPP by CFT
5306 samples taken from cattle, deer, wild sheep, reindeer and buffalo.
The samples were analyzed with two CFT antigens Mycoplasma
mycoides subsp. mycoides provided by CBPP OIE Reference
Laboratories from LINV Lisbon-Portugal and ISZ G.Caporale Teramo-
Infectious and Zoonotic Deseases (Public Health) 87